Insulin Glargine ELISA

Insulin glargine, sold under the brand name LANTUS among others, is a long-acting insulin, used in the management of type I and type II diabetes. It was approved for medical use in the United States in 2000 and is on the WHO's List of Essential Medicines. 

Mechanism of action: Insulin glargine is produced by recombinant DNA technology. It is an analogue of human insulin made by replacing the asparagine residue at position A21 of the A-chain with glycine and adding two arginine to the C-terminus (positions B31 and 32) of the B-chain. The resulting protein is soluble at pH 4 and forms micro- precipitates at physiological pH 7.4, making the molecule more soluble at an acidic pH and less soluble at physiological pH. The isoelectric shift also allows for the subcutaneous injection of a clear solution. Small amounts of insulin glargine are slowly released from micro-precipitates giving the drug a long duration of action (up to 24 hours) and no pronounced peak concentration.

Principle of the assay: The method employs the quantitative sandwich enzyme immunoassay technique. In the first step, samples and standards are pipetted into microwells. If Insulin Glargine is present in the samples or standards, it will form complex with Anti-insulin Glargine antibody, which is pre-coated onto microwells. In the second step, anti-Insulin Glargine Biotin conjugate is pipetted and incubated. In the third step, HRP conjugate is pipetted and incubated. Free HRP conjugate will be removed by a washing step. In the fouth step, TMB substrate is added to microwells and color develops proportionally to the amount of Insulin Glargine present in the samples or standards. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.

Krishgen is of the only companies in the world to manufacture these ELISA commercially.

Kit features:

• highly specific antibody pairs for detection and coating, with proprietary stabilisers and blockers for optimal noise:signal ratio.
• High specificity for the target analyte
• Lot to Lot consistency
• This kit has been validated as per EMA/FDA guidelines in line with ICH Code for Harmonization of Biological Assays
• Validated for both plasma and serum samples